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Thread-based isotachophoresis for DNA extraction and purification from biological samples

Citation

Chen, L and Cabot, JM and Paull, B, Thread-based isotachophoresis for DNA extraction and purification from biological samples, Lab on A Chip, 2021, (13) pp. 2565-2573. ISSN 1473-0189 (2021) [Refereed Article]


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DOI: doi:10.1039/D1LC00179E

Abstract

A rapid, low-cost, and disposable microfluidic thread-based isotachophoresis method was developed for the purification and preconcentration of nucleic acids from biological samples, prior to their extraction and successful analysis using quantitative polymerase chain reaction (qPCR). This approach extracts and concentrates protein-free DNA from the terminating electrolyte buffer, via a continuous sampling approach, resulting in significant focussing of the extracted DNA upon a 6 cm length nylon thread. The platform was optimised using the preconcentration of a fluorescent dye, showing a 600-fold concentration capacity within <5 min. The system was then applied to the one-step extraction of lambda DNA an E. coli bacteriophage spiked into whole blood, exhibiting the exclusion of PCR inhibitors. The extraction efficiency from the thread material following concentration was consistent, between 94.4113.9%. The determination of lambda DNA in whole blood was achieved within a linear range of 1.01 105 fg μL−1 (202 106 copies per μL). This technique demonstrates great potential for the development of thread-based affordable analytical and diagnostic devices based upon DNA and RNA isolation.

Item Details

Item Type:Refereed Article
Keywords:Thread-based isotachophoresis, DNA extraction, DNA purification, DNA, separation science
Research Division:Chemical Sciences
Research Group:Analytical chemistry
Research Field:Separation science
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the chemical sciences
UTAS Author:Chen, L (Miss Liang Chen)
UTAS Author:Cabot, JM (Dr Joan Cabot Canyelles)
UTAS Author:Paull, B (Professor Brett Paull)
ID Code:151715
Year Published:2021
Funding Support:Australian Research Council (DP170102572)
Web of Science® Times Cited:4
Deposited By:Chemistry
Deposited On:2022-08-03
Last Modified:2022-08-03
Downloads:0

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